Characterization of a reference material for BCR-ABL (M-BCR) mRNA quantitation by real-time amplification assays: Towards new standards for gene expression measurements Academic Article uri icon

Overview

MeSH Major

  • Gene Expression Profiling
  • Polymerase Chain Reaction
  • Protein-Tyrosine Kinases
  • RNA, Messenger

abstract

  • Monitoring of BCR-ABL transcripts has become established practice in the management of chronic myeloid leukemia. However, nucleic acid amplification techniques are prone to variations which limit the reliability of real-time quantitative PCR (RQ-PCR) for clinical decision making, highlighting the need for standardization of assays and reporting of minimal residual disease (MRD) data. We evaluated a lyophilized preparation of a leukemic cell line (K562) as a potential quality control reagent. This was found to be relatively stable, yielding comparable respective levels of ABL, GUS and BCR-ABL transcripts as determined by RQ-PCR before and after accelerated degradation experiments as well as following 5 years storage at -20 degrees C. Vials of freeze-dried cells were sent at ambient temperature to 22 laboratories on four continents, with RQ-PCR analyses detecting BCR-ABL transcripts at levels comparable to those observed in primary patient samples. Our results suggest that freeze-dried cells can be used as quality control reagents with a range of analytical instrumentations and could enable the development of urgently needed international standards simulating clinically relevant levels of MRD.

publication date

  • July 2007

Research

keywords

  • Academic Article

Identity

Language

  • eng

Digital Object Identifier (DOI)

  • 10.1038/sj.leu.2404716

PubMed ID

  • 17476280

Additional Document Info

start page

  • 1481

end page

  • 7

volume

  • 21

number

  • 7